Quantitative image analysis has become an indispensable tool for biologists using microscopy throughout basic biological and biomedical research. As a National Biomedical Technology Research Resource, our purpose is to develop and disseminate novel technologies throughout the biomedical research community.ĬOBA’s mission is to serve the cell biology community’s growing need for sophisticated software for light microscopy image analysis. (I) Differentiated adipocytes were overlaid with corresponding nuclei using the “ OverlayObjects” module.The Center for Open Bioimage Analysis (COBA) was established in 2020 with a P41 grant from the NIH National Institute of General Medical Sciences. (H) Objects below the area threshold were excluded from the final analysis using the “ FilterObjects” module. (G) Adipocytes were identified by size, shape, and intensity from the binary image using the “ IdentifyPrimaryObjects” module. (F) Identified objects from the “ SplitOrMergeObjects” module were converted to a binary image using the “ ConvertObjectsToImage” module. (E) Touching lipid droplets were grouped together using the “ SplitOrMergeObjects” module. (D) Identified lipid droplets using the “ IdentifyPrimaryObjects” and “ FilterObjects” modules. (C) Corresponding image of DAPI stained nuclei. (B) Conversion of original image to grayscale using the “ ColorToGray” module. Image also contains a magnified picture of a differentiated adipocyte to model the diversity in lipid droplet sizes within a single cell. (A) Original image of differentiated adipocytes with a high density of BODIPY stained lipid droplets. In conclusion, this novel image analysis tool can provide a more precise evaluation of lipid droplet and adipogenesis dysregulation, a critical need in the understanding of metabolic disorders.Īdipocyte image segmentation lipid droplet quantitative analyses. CellProfiler streamlines the lipid droplet phenotypic analysis of adipocytes compared to more traditional analysis methods. A clustering analysis is also possible using CellProfiler which allows for the quantification of total lipid content per individual adipocyte to provide insight into single-cell responsiveness to adipogenic stimuli. Our results show that CellProfiler is able to accurately identify a greater number of lipid droplets compared to ImageJ. For ImageJ, we used an already developed macro designed to identify particles and quantify their area, and for CellProfiler, we developed a new analysis pipeline. For the lipid droplet analysis, we used two approaches, the free online computer software of reference, ImageJ, and another free online computer software, CellProfiler. Therefore, the aims of this study were to develop an accurate, standardized approach to quantify lipid droplet size of mature adipocytes and a clustering approach to analyze the total lipid content per adipocyte. Nutrition, stress, or chemical exposure can dysregulate adipogenic differentiation and lipid metabolism. However, imaging tools for evaluating intracellular lipid droplets remain at their infancy. During differentiation, neutral lipids that accumulate in adipocytes can be detected using stains and used as an index of cell differentiation. Adipogenic differentiation is the process by which preadipocytes become mature adipocytes, cells that store energy and regulate metabolic homeostasis.
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